Quarternary ammonium compounds

ABSTRACT

Novel compounds and pharmaceutical compositions containing such compounds and possessing anti-tussive activity, and a method of administering the same to warm-blooded animals, including humans.

[0001] This application is a divisional of U.S. Ser. No. 10/144,607filed May 13, 2002, which is a divisional of U.S. Ser. No. 09/328,541filed Jun. 9, 1999, now U.S. Pat. No. 6,388,095 issued May 14, 2002,which claims priority of U.S. Provisional Application No. 60/088,587filed Jun. 9, 1998, all of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] Conventional cough preparations containing an effectiveanti-tussive agent such as codeine have long been used for thesymptomatic relief of coughs. However, codeine has various side effectswhich are undesirable.

[0003] Accordingly, the present invention relates to compounds andpharmaceutical compositions having anti-tussive activity, and a methodof treating warm-blooded animals affected by coughs by administering aneffective amount of the compounds or the pharmaceutical compositions ofthe invention.

SUMMARY OF THE INVENTION

[0004] The problems of the prior art have been overcome by the presentinvention, which provides pharmaceutical compositions possessinganti-tussive activity, and a method of administering the same towarm-blooded animals, including humans. The active anti-tussive agent inaccordance with the present invention is a novel quarternary ammoniumcompound represented by the following formula (I), or a solvate orpharmaceutically acceptable salt thereof:

[0005] wherein Y and E are independently selected from —CH₂-R₁₆ or agroup represented by the following formula (II):

[0006] wherein R, R₁, R₂, R₃, R₄, R₅, R₆ and R₁₆ are independentlyselected from hydrogen, C₁-C₈ alkyl, C₃-C₈ alkoxyalkyl and C₇-C₁₂aralkyl; m is an integer of from 1 to 8 and n is an integer of from 0 to8; A is selected from C₅-C₁₂ alkyl, a C₃-C₁₃ carbocyclic ring, and ringsystems selected from formulae (III), (IV), (V), (VI), (VII), (VIII),(IX) and (X):

[0007] where R₇, R₈, R₉ R₁₀, R₁₁ and R₁₂, are independently selectedfrom bromine, chlorine, fluorine, carboxy, hydrogen, hydroxy,hydroxymethyl, methanesulfonamido, nitro, sulfamyl, trifluoromethyl,C₂-C₇ alkanoyloxy, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₂-C₇ alkoxycarbonyl,C₁-C₆ thioalkyl, aryl and N(R₁₃,R₁₄) where R₁₃ and R₁₄ are independentlyselected from hydrogen, acetyl, methanesulfonyl and C₁-C₆ alkyl, and Zis selected from CH, CH₂, O, N and S, where Z may be directly bonded toX when Z is CH, or X may be a direct bond to Z when Z is N, or Z may bedirectly bonded to R₁₅ when Z is N and X is not a direct bond to Z, R₁₅is selected from hydrogen, C₁-C₆ alkyl, C₃-C₈ cyclocalkyl, aryl andbenzyl; and X is N—R₆ except when Z in A is nitrogen and X is a directbond to Z; An⁻ is the acid addition salt of a pharmaceuticallyacceptable acid or the anion from a pharmaceutically acceptable salt,and isolated enantiomeric, diastereomeric and geometric isomers thereof,and mixtures thereof, with the proviso that Y and E cannot both be—CH₂-R₁₆ in the same compound.

[0008] Within the respiratory tract the epithelium of the larynx,trachea, and larger bronchi contains sensory nerves that are responsiblefor cough. Coughing is initiated when sensory receptors in therespiratory tract receive stimuli of sufficient intensity to evoke anincrease in afferent nerve impulse activity. Cough reflexes can beprovoked easily by mechanical and chemical stimuli applied to theepithelium of either the larynx or tracheobronchial tree. There arethree main groups of airway sensory receptors which may be involved inthe cough reflex initiated from these sites: the slowly adapting stretchreceptors, the rapidly adapting (irritant) receptors (RARs), and thepulmonary and bronchial C-fibre receptors. Each is distributedthroughout the tracheobronchial tree and the last group is also presentin the alveolar wall. Irritant and C-fibre receptors have also beenidentified in the larynx. Stimulation of irritant receptors evokes thecough reflex; stimulation of the C-fibre receptors may either evoke orinhibit the cough reflex; stimulation of the slowly adapting stretchreceptors may facilitate the cough reflex.

[0009] Drugs which inhibit cough may act at a variety of sites, bothperipherally and centrally. For example, it is generally assumed thatthe anti-tussive effects of currently available opiates are mediatedcentrally through an action on the ‘cough centre’ in the medulla (D. T.Chou et al. J. Pharmacol. Exp. Ther. 1975, 194, 499). Thus the number ofpotential sites of action of anti-tussive drugs includes all componentsof the cough reflex pathway, from its initiation to its finalsynchronized motor response.

[0010] It has now been discovered that a class of cation channelmodulator/blockers are of potential use in the treatment and/orprevention of cough in warm-blooded animals including humans. The cationchannel modulator/blockers of the present invention inhibit theinitiation of an action potential by a cation channel at the peripheralfine afferent nerve ending or on peripheral sensory neuron that isresponsible for inducing cough or triggering cough reflex. The cationchannel at peripheral fine afferent nerve ending or on peripheralsensory neuron may be coupled as the transduction mechanism to one ormore sensory receptors described above (RARs, C-fibre receptors and theslowly adapting stretch receptors).

[0011] The cation channel at the peripheral fine afferent nerve endingor peripheral sensory neuron is identified to be a member of theacid-sensing ion channels (ASIC) or the dorsal root acid-sensing ionchannel (DRASIC). (see e.g. R. Waldmann et al. Curr. Opin. Neurobiol.1998, 8(3), 418).

[0012] In another embodiment, the present invention provides the cationchannel modulator/blockers that are quaternary ammonium compounds.

[0013] In another embodiment, the present invention provides the cationchannel modulator/blockers that are quaternary ammonium compounds offormula I.

[0014] In another embodiment, the present invention provides a methodfor the treatment and/or prevention of cough in warm-blooded animalsincluding human, which comprises administering to a warm-blooded animalin need thereof a therapeutically effective amount of an antitussivecomprising as the antitussive effective ingredient a modulator/blockerof a cation channel, or a pharmaceutically acceptable salt thereof.

[0015] In another embodiment, the present invention provides a methodfor the treatment and/or prevention of cough in warm-blooded animalsincluding human, which comprises administering to a warm-blooded animalin need thereof a therapeutically effective amount of an antitussivecomprising as the antitussive effective ingredient a modulator/blockerof a cation channel that is at the peripheral fine afferent nerve endingor on a peripheral sensory neuron of said warm-blooded animal, or apharmaceutically acceptable salt thereof.

[0016] In yet another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of a cation channel that is at the peripheral fineafferent nerve ending or on a peripheral sensory neuron which may becoupled as the transduction mechanism to one or more sensory receptorsof said warm-blooded animal, or a pharmaceutically acceptable saltthereof.

[0017] In still another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of a cation channel that inhibits initiation of anaction potential at the peripheral fine afferent nerve ending or on aperipheral sensory neuron of said warm-blooded animal, or apharmaceutically acceptable salt thereof.

[0018] In yet another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of a cation channel that inhibits initiation of anaction potential at the peripheral fine afferent nerve ending or on aperipheral sensory neuron that is responsible for inducing cough ortriggering cough reflex of said warm-blooded animal, or apharmaceutically acceptable salt thereof.

[0019] In still another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of a cation channel that inhibits from extracellularloci the initiation of an action potential at the peripheral fineafferent nerve ending or on a peripheral sensory neuron that isresponsible for inducing cough or triggering cough reflex of saidwarm-blooded animal, or a pharmaceutically acceptable salt thereof.

[0020] In still another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of an acid-sensing ion channel (ASIC), or apharmaceutically acceptable salt thereof.

[0021] In yet another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of a dorsal root acid-sensing ion channel (DRASIC), ora pharmaceutically acceptable salt thereof.

[0022] In still another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of an acid-sensing ion channel (ASIC) at theperipheral fine afferent nerve ending or on a peripheral sensory neuron,or a pharmaceutically acceptable salt thereof.

[0023] In still another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient amodulator/blocker of an acid-sensing ion channel (ASIC) at theperipheral fine afferent nerve ending or on a peripheral sensory neuronwhich may be coupled as the transduction mechanism to one or moresensory receptors, or a pharmaceutically acceptable salt thereof.

[0024] In yet another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient aquaternary ammonium compound functioning as a modulator/blocker of anacid-sensing ion channel (ASIC) at the peripheral fine afferent nerveending or on a peripheral sensory neuron, or a pharmaceuticallyacceptable salt thereof.

[0025] In yet another embodiment, the present invention provides amethod for the treatment and/or prevention of cough in warm-bloodedanimals including human, which comprises administering to a warm-bloodedanimal in need thereof a therapeutically effective amount of anantitussive comprising as the antitussive effective ingredient aquaternary ammonium compound of Formula (I) functioning as amodulator/blocker of an acid-sensing ion channel (ASIC) at theperipheral fine afferent nerve ending or on a peripheral sensory neuron,or a pharmaceutically acceptable salt thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0026]FIG. 1 is a flow diagram showing the layout of the experimentalapparatus used for cough determination; and

[0027]FIGS. 2A and 2B are expanded scale recordings of pressure changesderived from the differential pressure transducer during characteristicresponses exhibited by a guinea-pig during exposure to an aerosol ofcitric acid.

DETAILED DESCRIPTION OF THE INVENTION

[0028] As used herein, the following terms have the following meaning:

[0029] “Alkyl” refers to a branched or unbranched hydrocarbon fragmentcontaining the specified number of carbon atoms and having one point ofattachment. Examples include n-propyl A C₃ alkyl), isopropyl (also a C₃alkyl) and t-butyl (a C₄ alkyl).

[0030] “Alkoxyalkyl”, refers to an alkylene group substituted with analkoxy group. For example, methyoxyethyl (CH₃OCH₂CH₂—) and ethoxymethyl(CH₃CH₂OCH₂—) are both C₃ alkoxyalkyl groups.

[0031] “Alkylene” refers to a divalent radical which is a branched orunbranched hydrocarbon fragment containing the specified number ofcarbon atoms and having two points of attachment. An example ispropylene (—CH₂CH₂CH₂—), a C₃ alkylene.

[0032] “Aralkyl” refers to an alkylene group wherein one of the pointsof attachment is to an aryl group. An example is the benzyl group(C₆H₅CH₂—), a C₇ aralkyl group.

[0033] “Alkanoyloxy” refers to an ester substituent wherein the etheroxygen is the point of attachment to the molecule. Examples includepropanoyloxy (CH₃CH₂C(O)—O—), a C₃ alkanoyloxy and ethanoyloxy(CH₃C(O)—O), a C₂ alkanoyloxy.

[0034] “Alkoxy” refers to an O-atom substituted by an alkyl group, forexample methoxy (—OCH₃), a C₁ alkoxy.

[0035] “Alkoxycarbonyl” refers to an ester substituent wherein thecarbonyl carbon is the point of attachment to the molecule. Examplesinclude ethoxycarbonyl (CH₃CH₂(O)C═O), a C₃ alkoxycarbonyl, andmethoxycarbonyl (CH₃OC(O)—), a C₂ alkoxycarbonyl.

[0036] “Aryl” refers to aromatic groups which have at least one ringhaving a conjugated pi electron system and includes carbocyclic aryl,heterocyclic aryl (also known as heteroaryl groups) and biaryl groups,all of which may be optionally substituted. Carbocyclic aryl groups aregenerally preferred in the compounds of the present invention, whereinphenyl and naphthyl groups are preferred carbocyclic aryl groups.

[0037] “Cycloalkyl” refers to a ring, which may be saturated orunsaturated and monocyclic, bicyclic or tricyclic formed entirely fromcarbon atoms. An example is the cyclopentenyl group (C₅H₇—), which is afive carbon unsaturated cycloalkyl group.

[0038] “Carbocyclic” refers to a ring which may be either an aryl ringor a cycloalkyl ring, both as defined above.

[0039] “Thioalkyl” refers to a sulfur atom substituted by an alkylgroup, for example thiomethyl (CH₃S—), a C₁ thioalkyl.

[0040] The origin of the cough to be treated by the present invention isnot particularly limited, and can include virtually any respiratorydisorder, such as chronic obstructive pulmonary disease, tuberculosis,bronchitis, respiratory malignancies, asthma, allergy, pulmonaryfibrosis, respiratory tract inflammation, emphysema, pneumonia, lungcancer, presence of foreign bodies, soar throat, common cold, influenza,respiratory tract infection, bronchoconstriction, inhalation ofirritants, smoker's cough, chronic non-productive cough, neoplasticcough, cough due to angiotension converting enzyme (ACE) inhibitortherapy, etc. Cough may also occur without a known cause.

[0041] The compounds of the present invention are quaternary ammoniumsalts represented by the following formula (I), or a solvate orpharmaceutically acceptable salt thereof:

[0042] wherein Y and E are independently selected from —CH₂-R₁₆ or agroup represented by the following formula (II):

[0043] wherein R, R₁, R₂, R₃, R₄, R₅, R₆ and R₁₆ are independentlyselected from hydrogen, C₁-C₈ alkyl, C₃-C₈ alkoxyalkyl and C₇-C₁₂aralkyl; m is an integer of from 1 to 8 and n is an integer of from 0 to8; A is selected from C₅-C₁₂ alkyl, a C₃-C₁₃ carbocyclic ring, and ringsystems selected from formulae (III), (IV), (V), (VI), (VII), (VIII),(IX) and (X):

[0044] where R₇, R₈, R₉, R₁₀, R₁₁ and R₁₂ are independently selectedfrom bromine, chlorine, fluorine, carboxy, hydrogen, hydroxy,hydroxymethyl, methanesulfonamido, nitro, sulfamyl, trifluoromethyl,C₂-C₇ alkanoyloxy, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₂-C₇ alkoxycarbonyl,C₁-C₆ thioalkyl, aryl and N(R_(13,)R₁₄) where R₁₃ and R₁₄ areindependently selected from hydrogen, acetyl, methanesulfonyl and C₁-C₆alkyl, and Z is selected from CH, CH₂, O, N and S, where Z may bedirectly bonded to X when Z is CH, or X may be a direct bond to Z when Zis N, or Z may be directly bonded to R₁₅ when Z is N and X is not adirect bond to Z, R₁₅ is selected from hydrogen, C₁-C₆ alkyl, C₃-C₈cyclocalkyl, aryl and benzyl; and X is N—R₆ except when Z in A isnitrogen and X is a direct bond to Z; An⁻ is the acid addition salt of apharmaceutically acceptable acid or the anion from a pharmaceuticallyacceptable salt, and isolated enantiomeric, diastereomeric and geometricisomers thereof, and mixtures thereof, with the proviso that Y and Ecannot both be —CH₂-R₁₆ in the same compound.

[0045] A preferred compound of the present invention is a compound ofthe formula (I) as represented byN,N-Bis-[(1-Indolinyl)carbonymethyl]dimethylammonium chloride with thefollowing structure:

[0046] Another preferred compound of the present invention is a compoundof the formula (I) as represented byN-[(1-Indolinyl)carbonylmethyl]-N-(phenylcarbamoylmethyl)dimethylammoniumchloride with the following structure:

[0047] Another preferred compound of the present invention is a compoundof the formula (I) as represented byN-[(1-Indolinyl)carbonylmethyl]triethylammonium chloride with thefollowing structure:

[0048] Another preferred compound of the present invention is a compoundof the formula (I) as represented byN,N-Bis-[(1-Naphthyl)carbonylmethyl]dimethylammonium chloride with thefollowing structure:

[0049] The compounds of the present invention may be prepared by analogywith known synthetic methodology (see, e.g., Belgian Patent 614,154,which follows from Swedish Patent 1779/71, the disclosures of which areherein incorporated by reference). A conventional route of synthesisinvolves three steps and can be described (as in the aforementionedpatent, see also T. Takahashi, J. Okada, M. Hori, A. Kato, K. Kanematsu,and Y. Yamamoto, J. Pharm. Soc. Japan 76, 1180-6 (1956)) as follows.

[0050] In a first step an aromatic amine is reacted with chloracetylchloride in a suitable solvent such as dichloromethane and in thepresence of triethylamine. The reaction is conducted at low temperature(−15° C.) and the desired product is recovered from the reaction mixtureby conventional organic chemistry techniques, and if necessary, can bepurified by chromatography techniques. In a second step, the abovechlorinated derivative can be reacted with a tertiary amine in a solventsuch as methanol with a catalyst (e.g., potassium iodide) to form aquaternary ammonium salt. The chlorinated intermediate can react as wellwith a secondary amine to provide the corresponding tertiary amine,which is then further reacted with a chlorinated derivative to form aquaternary ammonium salt.

[0051] The synthetic procedures described herein, especially when takenwith the general knowledge in the art, provide sufficient guidance tothose of ordinary skill in the art to perform the synthesis, isolation,and purification of the compounds of the present invention.

[0052] Suitable pharmaceutically acceptable salts include acid additionsalts of acids such as hydrochloric, hydrobromic, benzenesulfonic(besylate), benzoic, camphorsulfonic, ethanesulfonic, fumaric, gluconic,glutamic, isethionic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, pamoic, pathothenic, succinic, p-toluenesulfonic, phosphoric,sulphuric, citric, tartaric, lactic and acetic acid, although thepreferred acid addition salt is the hydrochloride salt.

[0053] The magnitude of the therapeutic or prophylactic dose of thecompounds of the present invention in the treatment and/or prevention ofcough will depend upon the severity and nature of the condition beingtreated and the route of administration. The dose and the frequency ofthe dosing will also vary according to age, body weight and response ofthe individual patient. In general, the total daily dose range for thecompounds of the present invention for the treatment or prevention ofcough is from about 0.1 to about 800 mg in single or repeated doses.

[0054] Any suitable route of administration may be employed to providean effective dosage of the compounds of the present invention, althoughadministration by inhalation is preferred, most preferably in aerosolform. Suitable forms of administration include, but are not limited to,inhalation (delivered by, e.g., metered-dose inhaler, jet nebulizer,ultrasonic nebulizer, dry powder inhaler, etc.), nasal sprays,nebulization, oral administration such as via tablets, capsules,lozenges, syrups, sprays, suspensions, elixirs, gargles, and otherliquid preparations, aerosol foams, parental administration, andsublingal administration.

[0055] The compounds of the present invention can includepharmaceutically acceptable carriers and other conventional additives,including aqueous based carriers, co-solvents such as ethyl alcohol,propylene glycol and glycerin, fillers, lubricants, wetting agents,flavoring agents, coloring agents, emulsifying, suspending or dispersingagents, suspending agents, etc. For aerosol delivery of the compounds ofthe present invention, pharmaceutically acceptable diluents, carriers,and/or propellants may be included in the formulations for use inappropriate devices. These are prepared by procedures well known tothose skilled in the art (see e.g., Medication Teaching Manual, 5th Ed.,Bethesda, Md., American Society of Hospital Pharmacists, 1991).

[0056] The compositions of the present invention may optionally includeother known therapeutic agents, including decongestants such aspseudoephedrine HCl, phenylephrine HCl and ephedrine HCl, non-steroidalanti-inflammatory drugs such as acetaminophen, aspirin, phenacetin,ibuprofen and ketoprofen, expectorants such as glyceryl guaiacolate,terpin hydrate and ammonium chloride, antihistamines such aschlorpheniramine maleate, doxylamine succinate, brompheniramine maleateand diphenhydramine hydrochloride, and anesthetic compounds such asphenol.

[0057] The following examples are offered by way of illustration and notby way of limitation.

EXAMPLE 1 N,N-Bis-[(1-Indolinyl)carbonylmethyl]dimethylammonium chloridei) Chloroacetindolide

[0058] To a chilled (−13° C.) solution of indoline (12 g, 100 mmol) andtriethylamine (37 ml, 262 mmol) in dichloromethane (350 ml) was addeddropwise a solution of chloroactyl chloride (10.6 ml, 131 mmol) indichloromethane (250 ml). The reaction mixture was stirred at −13° C.for another 30 minutes and then allowed to warm up to room temperature.The reaction mixture was washed with water (600 ml), the aqueous layerwas collected and extracted once more with dichloromethane (200 ml). Thecombined organic layers were dried over sodium sulfate and the solventwas evaporated in vacuo to yield the crude title compound. Purificationby dry column chromatography with mixtures of ethyl acetate-hexanes aseluents followed by recrystallization from methanol provided 16 g of thetitle compound.

ii) Dimethylaminoacetindolide

[0059] A mixture of chloroacetindolide (8.5 g, 43.4 mmol) indimethylamine 40% wt. solution in water (85 ml) was refluxed for onehour. The cooled reaction mixture was partitioned between 1M NaOHaqueous solution (85 ml) and dichloromethane (170 ml). The aqueous layerwas collected and extracted once more with dichloromethane (85 ml). Thecombined organic extracts were back-washed with water 9200 ml, 100 ml)and dried over sodium sulfate. Evaporation of the solvent in vacuoyielded 8.8 g (95% yield) of the title compound.

iii) N,N-Bis-[(1-Indolinyl)carbonylmethyl]dimethylammonium chloride

[0060] A mixture of chloroacetindolide (1.96 g, 10 mmol),dimethylaminoacetindolide (2.04 g, 10 mmol) and potassium iodide (0.017g, 0.1 mmol) in methanol (20 ml) was heated to 80° C. for 16 hours. Tothe cooled reaction mixture was added diethyl ether (20 ml). Theresulting precipitate was collected and recrystallized from a mixture ofmethanol-diethyl ether (1:1, v/v) to yield 3.35 g (84% yield) of thetitle compound, characterized by elemental analysis as set forth inTable 1.

EXAMPLE 2

[0061]N-[(1-Indolinyl)carbonylmethyl]-N-(phenylcarbamoylmethyl)dimethylammoniumchloride

i) Chloroacetindolide

[0062]

[0063] To a chilled (−13° C) solution of aniline (18.2 g, 200 mmol) andtriethylamine (73 ml, 520 mmol) in dichloromethane (350 ml) was addeddropwise a solution of chloroactyl chloride (21.1 ml, 260 mmol) indichloromethane (250 ml). The reaction mixture was stirred at −13° C.for another 30 minutes and then allowed to warm up to room temperature.The reaction mixture was washed with water (600 ml), the aqueous layerwas collected and extracted once more with dichloromethane (200 ml). Thecombined organic layers were dried over sodium sulfate and the solventwas evaporated in vacuo to yield the crude title compound. Purificationby dry column chromatography with mixtures of ethyl acetate-hexanes aseluents followed by recrystallization from methanol provided 29.6 g (87%yield) of the title compound.

ii) Dimethylaminoacetindolide

[0064] See steps i and ii of Example 1.

iii)N-[(1-Indolinyl)carbonylmethyl]-N-(phenylcarbamoylmethyl)dimethylammoniumchloride

[0065] A mixture of chloroacetindolide (1.20 g, 7.08 mmol),dimethylaminoacetindolide (1.44 g, 7.08 mmol) and potassium iodide(0.012 g, 0.071 mmol) in methanol (15 ml) was heated to 80° C. for 16hours. To the cooled reaction mixture was added diethyl ether (15 ml).The resulting precipitate was collected and recrystallized from amixture of methanol-diethyl ether (1:1, v/v) to yield 2.18 g (82% yield)of the title compound, characterized by elemental analysis as set forthin Table 1.

EXAMPLE 3 N-[(1-Indolinyl)carbonylmethyl]triethylammonium chloride I)Chloroacetindolide

[0066] See step i of Example 1.

ii) N-[(1-Indolinyl)carbonylmethyl]triethylammonium chloride

[0067] A mixture of chloroacetindolide (1.8 g, 9.2 mmol), triethylamine(4.0 ml, 28.7 mmol) and potassium iodide (0.015 g, 0.092 mmol) inmethanol (20 ml) was heated to 80° C. for 36 hours. To the cooledreaction mixture was added diethyl ether (50 ml). The resultingprecipitate was collected and recrystallized from a mixture ofmethanol-acetone-diethyl ether (1:3:8, v/v/v) to yield 2.07 g (76%yield) of the title compound, characterized by elemental analysis as setforth in Table 1.

EXAMPLE 4 N,N-Bis-[(1-Naphthyl)carbonylmethyl]dimethylammonium chloridei) Chloroacetindolide

[0068] To a chilled (0° C.) solution of 1-naphthylamine (25 g, 174 mmol)and potassium carbonate (29 g, 209 mmol) in chloroform (150 ml) wasadded a solution of chloroactyl chloride (15.3 ml, 192 mmol) inchloroform (100 ml). The solid was collected, washed with water (300ml), and then extracted with diethyl ether via Soxhlet extraction toyield 17.5 g (46% yield) of the title compound.

ii) Dimethylaminoacetyl-1-naphthylamide

[0069] A mixture of chloroacetyl-1-naphthylamide (8.96 g, 39 mmol) indimethylamine 40% wt. solution in water (100 ml) was refluxed for onehour. The cooled reaction mixture was partitioned between 1M NaOHaqueous solution (80 ml) and dichloromethane (100 ml). The aqueous layerwas collected and extracted once more with dichloromethane (100 ml). Thecombined organic extracts were back-washed with water (2×50 ml) anddried over sodium sulfate. Evaporation of the solvent in vacuo yielded8.87 g (100% yield) of the title compound.

iii) N,N-[(1-Naphthyl)carbonylmethyl]dimethylammonium chloride

[0070] A mixture of chloroacetyl-l-naphthylamide (6.59 g, 30 mmol),dimethylaminoacetyl-1-naphthylamide (6.85 g, 30 mmol) and potassiumiodide (50 mg, 0.3 mmol) in anhydrous m-xylene (30 ml) was refluxed for1 hour. The resulting precipitate was collected while the reactionmixture was still hot and was washed with hot portions of ethyl alcoholto yield 4.6 g (34% yield) of the title compound. TLC analysis andcapillary electrophoresis of an aliquot sample show the compound to bepure. TABLE 1 Example Calculated Found # Formula M.W. (%) (%) 1C₂₂H₂₆N₃O₂Cl 399.92 C 66.07 C 65.42 H 6.57 H 6.55 N 10.51 N 10.30 2C₂₀H₂₄N₃O₂Cl 373.88 C 64.25 C 63.91 H 6.39 H 6.47 N 11.24 N 11.13 3C₁₆H₂₅N₂OCl. 350.88 C 54.77 C 55.97 H 8.88 3H₂O H 8.90 N 7.98 N 8.09 4C₂₆H₂₆N₃O₂Cl 447.96 C 69.71 C 69.63 H 6.07 H 5.85 N 9.38 N 9.28

EXAMPLE 5

[0071] The Following method was one of the general methods available todetermine the antitussive activity of the compounds of the presentinvention.

[0072] Male albino Dunkin-Hartley strain guinea-pigs (weight 300-400 g)were supplied by Harlan UK Ltd., Bicester, Oxon, UK.

[0073] The method used was modified from that described by Adcock J.J.., Schneider C. and Smith T. W., “Effects of Morphine and a NovelOpioid Pentapeptide BW443C, on Cough, Nociception and Ventilation in theUnanaesthetized Guinea-pig”, Br. J. Pharmacol., 93, 93-100 (1988).Individual conscious guinea-pigs were placed unrestrained into a sealedpurpose built perspex exposure chamber (3,000 cm³ volume) and allowed toacclimatize prior to aerosol administration. The layout of theexperimental apparatus used is shown in FIG. 1.

[0074] Cylinder air was introduced into the exposure chamber at a flowrate of 1 liter/min, maintained by a needle valve and monitored by arotameter. From the rotameter the air passed through the cup of anultrasonic nebulizer (DeVilbis UltraNeb 2000) which was used to generateaerosols of drug or citric acid at 0.15 ml/min. A Fleischpneumotachograph, connected to a differential pressure transducer (Grassmodel PT5) was attached to the outflow from the exposure chamber andprovided a measurement of airflow from the chamber. The differentialpressure transducer was connected to a Grass polygraph from which a hardcopy record was produced. The output from the polygraph was directed toa computerized data acquisition system (Poh-Ne-Mah) for real timerecording of data. A tie-clip microphone was placed in the exposurechamber and connected via a pre-amplifier to a loudspeaker output toprovide the observer with an audio monitor of responses.

[0075] Cough responses were induced by exposure to an aerosol of citricacid (1M) for 10 minutes. Animals were continuously monitored by trainedobserver, and the number of coughs were counted during a 15 minuteperiod from commencement of the citric acid aerosol administration.Three characteristic responses were produced by exposure to citric acid:cough, sneeze and “wet dog” shake.

[0076] The three types of response were differentiated primarily bysound and visual observation. Confirmation of the numbers of multiplecoughs was determined by reference to the change in flow rate displayedby the Poh-Ne-Mah system monitor. Printouts demonstrating the pressurechanges characteristic of the different response to irritant are shownin FIGS. 2A and 2B. Data recorded for individual guinea-pigs on thePoh-Ne-Mah system was stored on an optical disk. Each cough was markedon the Grass polygraph paper trace, and from these record numbers,frequency and time of onset of coughs were determined. The coughresponse was defined by a characteristic coughing sound and behavior,associated with a marked biphasic pressure change. The biphasic pressurechanges associated with a sneeze were not of as great a magnitude asthose associated with a cough, the secondary rise in pressure also beingfar less than during a cough (FIG. 2B). The sound of a sneeze differedfrom that of a cough, and sneezing was associated with nose rubbingactivity. The third response, a “wet dog” shake, produced a rise inpressure only (FIG. 2A) and lacked the definitive sound of a cough orsneeze.

[0077] Quantities of drugs were weighed out and dissolved in a vehicle.Equal volumes were aliquotted into sample tubes before being passed,together with another sample tube containing the same volume of vehicle,to an independent observer for coding. Pre-treatments were matched byconcentration together with a vehicle control group. Five guinea-pigswere randomly allocated to each treatment group. Animals werepre-treated with either vehicle (0.9% sterile saline), lidocaine or testdrugs for 5 minutes immediately prior to citric acid aerosol exposure.Test drugs and lidocaine were administered as aerosols at concentrationsof 0.1, 1.0 and 10.0 mg/ml. The sequence of pre-treatment administrationwas determined according to a 4×4 Latin Square design.

[0078] Data presented as the mean ±SEM number of coughs produced byindividual guinea-pigs within each group during the 15 minuteobservation period or mean±SEM latency of cough were analyzed using oneway analysis of variance to compare mean responses between matchedgroups of animals (doses) and between unmatched groups (treatments)followed by the Tukey-Kramer multiple comparison test where appropriate.

[0079] In one set of experiments using the general protocol describedabove, the antitussive activity ofN-[(1-Indolinyl)carbonylmethyl]-N-(phenylcarbamoylmethyl)dimethylammoniumchloride (Compound2) was tested. Results showed that pre-treatment ofguinea pigs with aerosols of Compound 2 at 10.0 mg/ml immediately beforeexposure to citric acid (1M) inhibited cough responses by 56% comparedwith vehicle (distilled water) pre-treated guinea pigs over the 15minute observation period. At the same time the mean latency of coughonset was prolonged by four fold.

[0080] The antitussive activity ofN-[(1-Indolinyl)carbonyl-methyl]triethylammonium chloride (Compound 3)was similarly tested. Pre-treatment of guinea pigs with aerosols ofCompound 3 at 10.0 mg/ml immediately before exposure to citric acid (1M)reduced cough responses by 36% compared with vehicle (distilled water)pre-treated guinea pigs over the 15 minute observation period. Resultsfrom the same experiment showed the time to the first recorded coughresponse elicited by citric acid was prolonged by Compound 3pre-treatment immediately prior to citric acid exposure, when latency ofcough onset increased 2.5 fold.

EXAMPLE 6

[0081] In another experiment similar to that described above in Example5, the duration of the antitussive effects of the compounds of thepresent invention against citric acid-induced cough responses wereinvestigated in conscious guinea pigs. Test agents or vehicle wereadministered as aerosol pre-treatments (10 mg/ml, 5 minute duration) at5 minutes, 30 minutes, 1 hour, 2 hours and 4 hours prior to induction ofcough responses by citric acid aerosol.

EXAMPLE 7

[0082] The antitussive effects of a 5 minute pre-treatment withaerosolized compounds of the present invention and lidocaine oncapsaicin aerosol-induced cough were investigated in consciousguinea-pigs using a method similar to that described in Example 5.

EXAMPLE 8

[0083] Therapeutic treatment with the compounds of the present inventionalso can be determined by a similar method as described in Example 5.The antitussive effects of compounds of the present invention andlidocaine administered after induction of cough responses by exposure tocitric acid aerosol were investigated in conscious guinea pigs. Vehicleor test agents were administered as aerosols (10 mg/ml; 5 minuteduration) 2 minutes after exposure to citric acid aerosol began. Coughresponses were recorded during a 15 minute observation period (t=0 tot=15 minutes) from initiation of the citric acid exposure.

EXAMPLE 9 Investigation of Antitussive Activity of Aerosolized TestCompound on Citric Acid-Induced Cough Responses in Conscious Rabbits

[0084] Protocol

[0085] Twenty-two male New Zealand white rabbits were randomly allocatedto either of two groups of 11 rabbits.

[0086] Pairs of rabbits (control versus test) were placed in individualexposure chambers with an airflow of 5 liter/min through the chambers.

[0087] Each rabbit was exposed to ozone (3 ppm) for 1 hour.

[0088] The rabbits were then immediately exposed to aerosols of eithervehicle (chamber 1) or test compound (10 mg/ml, chamber 2) at anebulization rate of 0.9 ml/min.

[0089] Cough responses were induced with citric acid aerosol (1.6 M)Coughs were counted during the 10 minute exposure to citric acid.

[0090] All rabbits were exposed to ozone before vehicle or test drugpre-treatment.

[0091] All publications and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication or patent application was specificallyand individually incorporated by reference.

[0092] From the foregoing, it will be appreciated that, althoughspecific embodiments of the invention have been described herein forpurposes of illustration, various modifications may be made withoutdeviating from the spirit and scope of the invention. Accordingly, theinvention is not limited except as by the appended claims.

What is claimed is:
 1. A method for the treatment and/or prevention of cough in warm-blooded animals including human, which comprises administering to a warm-blooded animal in need thereof a therapeutically effective amount of a compound of the following formula (I) or a solvate of pharmaceutically acceptable salt thereof:

wherein Y and E are independently selected from CH₂-R₁₆ or a group represented by the following formula (II):

wherein R, R₁, R₂, R₃, R₄, R₅, and R₁₆ are independently selected from hydrogen, C₁-C₈ alkyl, C₃-C₈ alkoxyalkyl and C₇-C₁₂ aralkyl; m is an integer of from 1 to 8 and n is an integer of from 0 to 8; A is selected from C₅-C₁₂ alkyl, a C₃-C₁₃ carbocyclic ring, and ring systems selected from formulae (III), (IV), (V), (VI), (VII), (VIII), (IX) and (X):

where R₇, R₈, R₉, R₁₀, R₁₁ and R₁₂ are independently selected from bromine, chlorine, fluorine, carboxy, hydrogen, hydroxy, hydroxymethyl, methanesulfonamido, nitro, sulfamyl, trifluoromethyl, C₂-C₇ alkanoyloxy, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₂ C₇ alkoxycarbonyl, C₁-C₆ thioalkyl, aryl and N(R₁₃, R₁₄) where R₁₃ and R₁₄ are independently selected from hydrogen, acetyl, methanesulfonyl and C₁-C₆ alkyl, and Z is selected from CH, CH₂, O, N and S, where Z may be directly bonded to X when Z is CH, or X may be a direct bond to Z when Z is N, or Z may be directly bonded to R₁₅ when Z is N and X is not a direct bond to Z, R₁₅ is selected from hydrogen, C₁-C₆ alkyl, C₃-C₈ cyclocalkyl, aryl and benzyl; and X is N—R₆ except when Z in A is nitrogen and X is a direct bond to Z; An⁻ is the acid addition salt of a pharmaceutically acceptable acid or the anion from a pharmaceutically acceptable salt, and isolated enantiomeric, diastereomeric and geometric isomers thereof, and mixtures thereof, with the proviso that Y and E cannot both be —CH₂-R₁₆ in the same compound, or a pharmaceutical composition comprising an effective amount of a compound of formula (I) and a pharmaceutically acceptable carrier, diluent or excipient.
 2. The method of claim 1, wherein said compound is N,N-Bis-[(1-Indolinyl)carbonylmethyl]dimethylammonium chloride.
 3. The method of claim 1, wherein said compound is N-[(1-Indolinyl)carbonylmethyl]-N-(phenylcarbamoylmethyl)dimethylammonium chloride.
 4. The method of claim 1, wherein said compound is N-[(1-Indolinyl)carbonylmethyl]triethylammonium chloride.
 5. The method of claim 1, wherein said compound is N,N-Bis-[(1-Naphthyl)carbonylmethyl])dimethylammonium chloride.
 6. A method for the treatment and/or prevention of cough in warm-blooded animals including human, which comprises administering to a warm-blooded animal in need thereof a therapeutically effective amount of an anti-tussive comprising as the anti-tussive effective ingredient a modulator/blocker of cation channel, or a pharmaceutically acceptable salt therof.
 7. A method according to claim 6, wherein said cation channel is at peripheral fine afferent nerve ending or on peripheral sensory neuron of said warm-blooded animal.
 8. A method according to claim 7, wherein said cation channel at peripheral fine afferent nerve ending or on peripheral sensory neuron is present in the respiratory tract of said warm-blooded animal.
 9. A method according to claim 7, wherein said cation channel at peripheral fine afferent nerve ending or on peripheral sensory neuron may be coupled as the transduction mechanism to one or more sensory receptors in the respiratory tract.
 10. A method according to claim 9, wherein said sensory receptors comprise the rapidly adapting (irritant) receptors (RARs), the slowly adapting stretch receptors, and the pulmonary and bronchial C-fibre receptors.
 11. A method according to claim 7, wherein said modulator/blocker inhibits initiation of an action potential at the peripheral fine afferent nerve ending or peripheral sensory neuron.
 12. A method according to claim 7, wherein said modulator/blocker inhibits initiation of an action potential generated by activation of one or more sensory receptors according to claim 9 at the peripheral fine afferent nerve ending or on peripheral sensory neuron.
 13. A method according to claim 7, wherein said modulator/blocker inhibits initiation of an action potential at the peripheral fine afferent nerve ending or peripheral sensory neuron that is responsible for inducing cough or triggering cough reflex.
 14. A method according to claim 7, wherein said modulator/blocker inhibits from extracellular loci the initiation of an action potential at the peripheral fine afferent nerve ending or on peripheral sensory neuron that is responsible for inducing cough or triggering cough reflex.
 15. A method according to claim 7, wherein said cation channel at peripheral fine afferent nerve ending or on peripheral sensory neuron is an acid-sensing ion channel (ASIC).
 16. A method according to claim 7, wherein said cation channel at peripheral fine afferent nerve ending or on peripheral sensory neuron is a dorsal root acid-sensing ion channel (DRASIC).
 17. A method according to claim 6, wherein said modulator/blocker is a quaternary ammonium compound, or a pharmaceutically acceptable salt thereof.
 18. A method according to claim 6, wherein said modulator/blocker is a quaternary ammonium compound of the following Formula (I), or a pharmaceutically acceptable salt thereof:

wherein Y and E are independently selected from CH₂-R₁₆ or a group represented by the following formula (II):

wherein R, R₁, R₂, R₃, R₄, R₅ and R₁₆ are independently selected from hydrogen, C₁-C₈ alkyl, C₃-C₈ alkoxyalkyl and C₇-C₁₂ aralkyl; m is an integer of from 1 to 8 and n is an integer of from 0 to 8; A is selected from C₅-C₁₂ alkyl, a C₃-C₁₃ carbocyclic ring, and ring systems selected from formulae (III), (IV), (V), (VI), (VII), (VIII), (IX) and (X):

where R₇, R₈ R₉, R₁₀, R₁₁ and R₁₂ are independently selected from bromine, chlorine, fluorine, carboxy, hydrogen, hydroxy, hydroxymethyl, methanesulfonamido, nitro, sulfamyl, trifluoromethyl, C₂-C₇ alkanoyloxy, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₂-C₇ alkoxycarbonyl, C₁-C₆ thioalkyl, aryl and N(R₁₃, R₁₄) where R₁₃ and R₁₄ are independently selected from hydrogen, acetyl, methanesulfonyl and C₁-C₆ alkyl, and Z is selected from CH, CH₂, O, N and S, where Z may be directly bonded to X when Z is CH, or X may be a direct bond to Z when Z is N, or Z may be directly bonded to R₁₅ when Z is N and X is not a direct bond to Z, R₁₅ is selected from hydrogen, C₁-C₆ alkyl, C₃-C₈ cyclocalkyl, aryl and benzyl; and X is N—R₆ except when Z in A is nitrogen and X is a direct bond to Z; An⁻ is the acid addition salt of a pharmaceutically acceptable acid or the anion from a pharmaceutically acceptable salt, and isolated enantiomeric, diastereomeric and geometric isomers thereof, and mixtures thereof, with the proviso that Y and E cannot both be —CH₂-R₁₆ in the same compound. 